GSM7658434: Human_Fibroblast_IgG; Homo sapiens; OTHER
Nuclei were isolated from cell populations by incubating cell pellets with a nuclear extraction hypotonic buffer (20 mM HEPES-KOH, pH 7.9, 10 mM KCl, 0.5mM spermidine, 0.1% Triton X-100, 20% glycerol, freshly added protease inhibitors) for 10 minutes on ice. Samples were centrifuged and nuclei were flash frozen. CUT&RUN enriched fragments isolated were used as input for a library build consisting of end repair and adenylation, NEBNext stem-loop adapter ligation, and subsequent purification with AMPure XP beads (Agencourt). Barcoded fragments were then amplified by 17 cycles of high-fidelity PCR and purified using AMPure XP.